Nextgen Dissection of the Genomic Basis of Kidney Development

描述(由申请人提供)：该提案的长期目标是进行基于NexGen RNA-Seq的对驱动肾脏发育的基因表达程序的剖析。中心思想很直截了当。我们建议以一种敏感和定量的方式定义在发育中的肾脏的每一种关键细胞类型中活跃的所有基因集。将编制一份特征全面的地图集，其中包括微小核糖核酸以及更正统的编码信使核糖核酸，并提供对替代核糖核酸加工事件的详细分析。最终的结果将是一个重要的资源，描述了定义和驱动发育中的肾脏每个部分的类似基因表达密码。该资源将确定在每种细胞类型中表达的所有生长因子、受体和转录因子。它将建立一套新的细胞类型特异性分子标记，用于突变分析。此外，它还将为突变或疾病引起的基因表达模式变化的全球分析提供一个标准基线。此外，我们建议使用芯片序列方法来发现转录因子Hoxa11、Lhx1和SIX2的直接下游靶标。它们都被证明在肾脏发育中发挥重要作用，通过确定它们的直接下游靶点，我们将更好地了解它们在肾脏形成过程中的确切作用。 与公共健康相关：这项拟议的工作将使用最新的革命性测序技术来表征发育中的肾脏许多不同细胞类型中的所有基因活动。这将使我们更深入地了解驱动肾脏形成过程的遗传程序，这反过来将帮助我们更好地了解如何修复因疾病而受损的肾脏。

DESCRIPTION (provided by applicant): The long-term objective of this proposal is to conduct a NexGen RNA-Seq based dissection of the gene expression program driving kidney development. The central thesis is straightforward. We propose to define, in a sensitive and quantitative manner, all of the sets of genes active in each critical cell type of the developing kidney. An atlas will be generated, comprehensive in character, including microRNAs as well as more orthodox coding mRNAs, and providing detailed analyses of alternative RNA processing events. The end result will be an essential resource, describing the analog gene expression code that defines and drives each compartment of the developing kidney. This resource will identify all of the growth factors, receptors and transcription factors expressed in each cell type. It will establish a set of novel cell type specific molecular markers, useful in mutant analyses. In addition it will provide a standard baseline for the global analysis of changes in gene expression patterns occurring as a result of mutation or disease. In addition we propose to use a Chip-Seq approach to discover the direct downstream targets of the transcription factors Hoxa11, Lhx1 and Six2. Each has been shown to play an important role in kidney development, and by identifying their direct downstream targets we will gain a better view of their precise roles in the process of nephrogenesis. PUBLIC HEALTH RELEVANCE: The proposed work would use the latest revolutionary sequencing technology to characterize all of the gene activities in the many different cell types of the developing kidney. This would lead to a much deeper understanding of the genetic program that drives the process of making a kidney, which in turn will in time help us better understand how to repair kidneys that have been damaged by disease.